This research continues our studies aimed at generating cytotoxic lymphocytes in vitro capable of lysing autologous human leukemia cells by culturing remission lymphocytes with (1)\X-irradiated autologous leukemia cells and X-irradiated allogeneic normal cells or with (2)\X-irradiated lymphocytes pooled from 20 unrelated normal individuals. Highly purified interferon (IF) and nontoxic polyribonucleotide inducers of IF also will be tested for their ability to activate patients' remission lymphocytes to lyse autologous malignant cells. Effector cells will be characterized with regard to expression of cell surface antigens by monoclonal antibodies, which we have found distinguish cytotoxic T lymphocytes (CTLs) from natural killer (NK) and NK-like cells. In addition, we intend to continue our studies of human NK and CTL-mediated lysis of autologous cells infected with herpes simplex virus that can produce severe infections in leukemia and lymphoma patients. We have found that conventional NK cells lyse autologous HSV-infected cells. We have begun cloning CTL against HSV-infected cells and are studying the nature of the HLA restriction and the virus-type specificity of the clones. We have found human CTL clones generated by stimulation with HSV-1 lyse autologous HSV-infected lymphoblastoid cell lines (LCLs). Results of experiments in which a panel of HLA-typed LCLs was used indicated that our HSV-1-specific CTL clones are restricted by HLA class II MB or DR antigens. Monoclonal antibodies to MR or to a DR framework determinant blocked cytotoxicity by these CTL clones. All CTL clones were found to proliferate when stimulated with HSV-1, and the proliferative response was restricted by the same HLA class II antigens as those that restricted cytotoxicity. Following HSV stimulation of the clones, interleukin-2 activity was detected in supernatants, thereby strongly suggesting that these clones are helper independent, cytotoxic T-cell clones (HITc). We have generated CTL clones to both HSV-1 and HSV-2 and have obtained both HSV-1 or HSV-2 type specific clones and HSV type common clones. The HSV specificity of the proliferative response was the same as that of the cytotoxic response. These results suggest that the HITc clones recognize the same HSV antigens and HLA antigens for both cytotoxic and proliferative activities. Studies are being continued to attempt to elucidate which viral glycoproteins may serve as recognition structures for the HITc clones. (IT)